ghenipus at my-deja.com wrote:
>> We are attempting to extract total RNA from
> soybean roots, epicotyls, hypocotyls, and
> cotyledons, with the eventual intent of running
> RT-PCR and Northern analysis. We've tried twice
> now with the Chomczynski & Sacchi procedure
> (flash-freeze in liquid Nitrogen, GTC, phenol/
> Sent via Deja.com http://www.deja.com/> Before you buy.
The procedure you outline should work very well for plant tissues.
In labs where I have worked the SDS-Phenol procedure in Ausubel works
great too and Trizol works great. you need to take care to freeze the
tissues in liquid N2, pulverize them with mortor and pestle, (keeping
frozen, and we always used a tissuemizer to ensure rapid mixing of the
denaturing agents. There is no need to RNAse out or DEPC treat any
thing. RNases simply dont get into those disposable plastics unless
someone contaminates them. Simply use unopened bags of tips, and
eppendorf tubes. Also redissolve the total Rna in formamide, not water.
You dont need column kits for total RNA.